Recovery of E.

coli O157:H7 from needle-tenderized raw and cooked beef loin steaks

Richard Holley, Kavitha Palaniappan and Roniele Cordeiro Department of Food Science University of Manitoba Winnipeg, MB R3T 2N2

A report prepared for CBC Marketplace October 24, 2012

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Samples of vacuum packed fresh boneless beef loins were purchased locally in Winnipeg by the CBC. In the first part of the work: - A concentration of 107 CFU/ml of E. coli O157:H7 was spread on the lean surface of the meat. Meat was given a single pass lean side up through a Hollymatic flat blade needle tenderizer. Needles entered the left side of steaks shown in Fig. 1 and did not penetrate the lower fat layer. - The meat was cut into four steaks, each about 4 cm thick. - Three steaks were analyzed raw and one steak was cooked at 71oC before analysis. - Each steak was cut into three samples as shown in Fig. 1.
Figure 1

Sample 1

Sample 2

Sample 3

Steaks were then sectioned horizontally in parallel with the surface. Two cuts were made horizontal to the inoculated surface from each sample at 1 cm intervals from the inoculated surface (located at the right in Fig. 2) to yield 3 portions (P) of each sample through the steaks from the lean to fat surface.

Figure 2 Sample 1 P3 P2 P1

Sample 2

Inoculated surface

Sample 3

1. Raw steak results Table 1: Enumeration of E. coli O157:H7 recovered from raw steaks (n=3) in duplicate. Position (P) 1 (surface) 2 (2 cm) 3 (3cm) Mean (Log CFU/g) 1 5.0 3.8 2.7 Mean Number (CFU/g) 100,000 6,310 501

CFU = colony forming units on cefixime tellurite sorbitol MacConkey (CTSMAC) agar.

2. Cooked steaks (71oC) - One steak was cooked until the middle region reached 71.1C using a Hamilton Beach electric grill. Temperature was monitored with a digital thermometer (DigiSense) equipped with a thermocouple inserted into the centre of the thickness of the steak from the side. - As with raw steaks, three horizontal portions were cut from each sample at 1cm intervals and analyzed in duplicate. Results from 71oC cooked steaks - No colonies were recovered on CTSMAC agar (specific for E. coli O157:H7) after 48h of incubation - E. coli was found on violet red bile (VRB) agar (specific for coliforms) after enrichment of cooked samples. 3. Cooked steaks (60oC) In the second part of the work: - two steaks were cut from a fresh loin, manually tenderized with a hand-held Jaccard meat tenderizer and cooked to an internal temperature of 60oC (Fig.3). Enumeration of E. coli O157:H7 25 g of each portion from each sample (n=6) of steak was added to 225 ml of peptone water. Diluted samples were plated in duplicate on CTSMAC and VRB agar using an automated spiral plating device. Plates were incubated at 37C for 48 h.

Figure 3

P3

P2

P1

Each cooked steak was cut into three samples and each was sectioned horizontally (parallel to the inoculated surface) into 1 cm portions from the lean (P1) to fat (P3) surface.
Sample 1

Results The concentration of E. coli O157:H7 on the meat surface was 5 Log CFU/ml before cooking. - Cooking steaks at 60C reduced E. coli O157:H7 by 3 Log CFU/ml depending on the region of the steak (Table 2). - No significant difference was found between the use of CTSMAC or VRB agar in the recovery of E. coli O157:H7 from cooked steaks - The presence of E. coli O157:H7 on CTSMAC and VRB agar was confirmed in all samples after enrichment of cooked samples for 24 hr. Table 2: Enumeration of E. coli O157:H7 (Log CFU/g and numbers/g) from 60oC cooked beef steaks Portion of steak CTSMAC VRB 1 (surface) 1.34 (22 cells) 1.74 (60 cells) 2 (middle) 0.22 (2 cells) 0.29 (2 cells) 3 (bottom) 0 0 Conclusions from raw and cooked steaks Mechanical tenderization translocated E. coli O157:H7 from the injected lean surface to the interior of treated meat to a depth of at least 3 cm.
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Cooking inoculated, mechanically tenderized steak at 71C (internal) eliminated E. coli O157:H7 from the meat when conventional methods for detection (plating) were used. Upon enrichment of these cooked samples viable E. coli were detected, but they would have been present at levels less than 0.2 cell/g (the limit of detection). Cooking inoculated mechanically tenderized steaks to an internal temperature of 60C left about 40 viable E. coli O157:H7 cells/g at the surface and 1 cm deep within the meat, as well as 2 cells/g to a depth of 2 cm. Clearly, this temperature was not as effective as 71C in eliminating E. coli O157:H7 from cooked steaks. While cooking steak internally contaminated with E. coli O157:H7 by mechanical tenderization to an internal temperature of 71oC gave assurance that the pathogen would be eliminated (gave a 5 log or decimal reduction), cooking inoculated and tenderized steak to 60oC yielded less than a 4 decimal reduction and therefore did not give assurance of its safety.